hygiena BAX-Q7 Molecular Pathogen Detection System Installation Guide

hygiena BAX-Q7 Molecular Pathogen Detection System

Product Information

Specifications

• Product Name: Yeast and Mold PCR Assay
• Action Levels: 10-50 cfu/g for pooled sample protocol, 25 cfu/g or above for non-pooled sample protocol
• Enrichment Replicates: 3 disrupter tubes for pooled sample protocol
• Sample Homogenate Dilution: 1:10 according to food type
• Incubation Time: 44 hours
• PCR Tablets Hydration Volume: 50 µl lysate from step 12

Product Usage Instructions

• Step 1: Sample Homogenization
• Homogenize the sample in a 1:10 dilution according to the food type.
• Step 2: Determine Sample Volume
• Determine the sample volume to be tested. Refer to the User Guide or table on the back of the reference card for guidance.
• Step 3: Transfer Sample
• Transfer the sample to a disrupter tube.
• Step 4: Incubate Disrupter Tubes
• Incubate the disrupter tubes for 44 hours.
• Step 5: Add DNA Stabilizer
• Add DNA stabilizer to the disrupter tubes.
• Step 6: Agitate in Disrupter Device
• Agitate the disrupter tubes in a disrupter device.
• Step 7: Create a Rack File
• Create a rack file for further processing.
• Step 8: Transfer the Lysis Reagent
• Transfer 12 mL of Protease YM lysis buffer to cluster tubes.
• Step 9: Transfer Disrupted Samples
• Transfer the disrupted samples from the disrupter tubes to the cluster tubes.
• Step 10: Heat Cluster Tubes
• Heat the cluster tubes for 20 seconds.
• Step 11: Cool Cluster Tubes
• Cool the cluster tubes in a cooling block.
• Step 12: PCR Tablet Hydration
• Hydrate PCR tablets with 50 µl lysate from step 12.
• Step 13: Initialize Cycler
• Initialize the cycler for PCR.
• Step 14: Arrange PCR Tubes
• A rrange PCR tubes in the cooling block.
• Step 15: Place Tubes in the Cycler
• Place the PCR tubes in the cycler and run the program.
• Step 16: Review Results

Unload the samples and review the results on the screen. Refer to the User Guide for detailed information on result interpretation.

FAQ:

• Q: What are the action levels for the pooled sample protocol?
• A: The action levels for the pooled sample protocol are 10-50 cfu/g.
• Q: Can the non-pooled sample protocol be used for action levels below 25 cfu/g?
• A: No, the non-pooled sample protocol is recommended for action levels of 25 cfu/g or above.

using instruction

1. Homogenize the sample in 1:10 dilution according to the food type.

2. Determine the sample volume to be tested.
   (See User Guide or table on the back of this reference card.)

3. Transfer the sample to the disrupter tube.
   Pooled sample protocol requires triplicate disrupter tubes.

4. Incubate disrupter tubes.

5. Add DNA stabilizer to disrupter tubes.
   DNA stabilizer

6. Agitate in disrupter device.

7. Create a rack file.

8. Add protease to YM lysis buffer.

9. Transfer lysis reagent made in step 8 to cluster tubes
   Lysis reagent mixture can be stored at 2 – 8 °C for up to one week

10. Transfer disrupted samples to cluster tubes.
    *Pooled sample protocol requires pooled volumes from disrupter tubes into 1 cluster tube.

11. Heat cluster tubes.

12. Cool cluster tubes in the cooling block.

• Steps 11 and 12 can also be performed using the Hygiena™ Automated Thermal Block. See the Automated Thermal Block User Guide for details and instructions. 13. Initialize cycler. 14. Arrange PCR tubes in the cooling block. 15. Hydrate PCR tablets with 50 μl lysate from step 12. 16. Place tubes in the cycler and run the program. 17. Unload samples and review results on the screen. See User Guide for details.

Pooled Sample Protocol

This ultra-sensitive protocol uses pooled samples from three disrupter tube enrichment replicates for action levels of 10-50 cfu/g.

If your action level is:

| Then transfer this volume of homogenate to 3 disrupter tubes:| And pool these volumes of disrupted samples for testing:
---|---|---
10 cfu/g| 400 µL| 7 µL from 3 replicates
20 cfu/g| 200 µL| 7 µL from 3 replicates
50 cfu/g| 80 µL| 7 µL from 3 replicates

Non-Pooled Sample Protocol

This protocol for yeast and mold testing can be used without pooling for action levels of 25 cfu/g or above.

If your action level is:

|

Then use this volume of homogenate:

---|---
25 cfu/g| 400 µL
50 cfu/g| 200 µL
100 cfu/g| 100 µL
500 cfu/g| 20 µL
1000 cfu/g| 10 µL

References

• Support | Hygiena

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